Drug Testing and Analysis最新文献

We report the misuse of a novel synthetic glucocorticoid, 9α-fluoro-6α-methylprednisolone (9F6MP) for the first time in camel racing and, to the best of our knowledge, human or other animal sports. During routine post-race drug testing of cameline plasma samples using liquid chromatography-tandem mass spectrometry we encountered an unknown peak with the same selected reaction monitoring traces as a dexamethasone formate adduct but at a different retention time. The product ion mass spectrum of the unknown peak in negative ion mode was identical to dexamethasone. However, significant differences were observed in positive ion mode. Based on mass spectral analysis, we postulated the unknown peak to be a 6-methyl-16-nor isomer of dexamethasone. Following the procurement of a commercial 9F6MP reference material, the unknown peak was successfully identified as this substance. Interestingly, previous research predicted a high potential for glucocorticoid and anti-inflammatory activity for 9F6MP. However, the therapeutic use of 9F6MP in camels has not been approved by any authorities, and any toxicities and side-effects potentially caused by 9F6MP have not been thoroughly evaluated. Therefore, the misuse of 9F6MP should be strictly controlled for the sake of animal welfare and the integrity of camel racing. The information described in this case report will be beneficial for other anti-doping laboratories in both human and animal sports for the purpose of doping control.

This study presents LC-HRMS/MS analyses of LGD-4033 post-administration urine samples, following hydrolysis with β-glucuronidase and liquid–liquid extraction, against chemically synthesized molecules that matched previously proposed metabolites, characterized by ¹H and ¹³C NMR. Using this targeted metabolic investigation approach and the direct comparison of retention times and mass spectral data (high-resolution full scan mass accuracy and collision-induced fragmentation patterns), in accordance with WADA's TD2023IDCR provisions, resulted in unambiguous structural elucidation of additional LGD-4033 metabolites, including (a) the epi-long-term dihydroxylated metabolite (M5a); (b) the epi-pyrrolidinone metabolite (M2d); (c) the (R,R)-diastereoisomer of the ring-opened hydroxylated metabolite (M4b); and (d) one of the two detected tris-hydroxylated metabolites (M6a). Additionally, a new, previously undescribed metabolite, which is a hydroxylated derivative of the pyrrolidinone metabolite M2c, was also detected up to 4 days post-administration and coded as M7. Metabolites M5a and M2d are detectable up to 21 days post-administration and can be considered additional long-term markers. These findings expand current knowledge of LGD-4033 metabolism. From a doping control perspective, the proposed synthetic pathways may facilitate the production of reference materials for the detection and identification of a more comprehensive metabolite profile that will increase metabolic certainty in future LGD-4033 adverse analytical findings.

For the past couple of years, black market products have appeared and were confirmed to contain genetic products coding for human erythropoietin (EPO). While being prohibited by the World Anti-Doping Agency (WADA), they could be used to produce endogenously more EPO hormone and hence increase performance. In a previous work, we demonstrated the potential of 20-μL dried blood spots (DBS) to detect the presence of EPO transgene in human blood down to 250 copies (12,500 copies/mL), despite lower sensitivity (30-fold) than in 1-mL fresh blood. As the use of DBS as a collection matrix for antidoping is going to expand in the near future, our aim was to develop and validate a new protocol to improve the sensitivity of gene doping detection from DBS. Three DBS devices were evaluated: polymeric Tasso-M20 (TASSO Inc.) and Mitra (Neoteryx), and cellulosic Protein Saver 903 (Whatman). The best results were achieved with polymeric DBS, and a full validation was performed for the detection of the EPO transgene using Tasso M-20 DBS; 1500 copies/mL were detected in 50% of cases and robust detection was obtained at 5000 copies/mL (100 copies transgene in 20-μL DBS) with the four spots of the Tasso device tested over several weeks. The results confirm that polymeric DBS can be used as an alternative to fresh blood for gene doping detection with high sensitivity simplifying also potential reanalysis in the future.

An innovative software tool for the rapid and efficient simulation of the metabolism of new psychoactive substances (NPS) was developed, based on the open-source project mzmine, and applied. NPS are compounds designed to mimic the psychotropic effects of established illicit drugs while circumventing drug legislation. These compounds are developed solely regarding their desired effects, thus possibly leading to harmful side effects including the formation of toxic metabolites. Analytical reference standards, needed to carry out metabolic studies, are not immediately available because emerging NPS are primarily discovered subsequent to drug confiscations. Using these confiscated substances in traditional metabolic in vivo or in vitro studies is often not possible due to the substances being impure or being a part of a mixture of different NPS. Therefore, a software tool was developed to streamline the evaluation of data acquired by the online combination of electrochemistry and mass spectrometry for the simulation of NPS metabolism. Using this tool, it is possible to generate mass voltammograms directly from mass spectrometric raw data. Combining this newly implemented tool with existing filtering algorithms in mzmine, we simulated the metabolism of the synthetic cannabinoid receptor agonist (SCRA) methyl 3,3-dimethyl-2-[1-(pent-4-en-1-yl)-1H-indazole-3-carboxamido] butanoate (MDMB-4en-PINACA) from a mixed solution of different NPS. Fragmentation data indicated that one of the transformation products found for MDMB-4en-PINACA is likely of a quinoid structure. The potential formation of this possibly highly reactive quinoid metabolite could be a first hint for possible causes of adverse side effects frequently reported after the recreational use of MDMB-4en-PINACA and related SCRAs.

New psychoactive substances (NPS), especially synthetic cannabinoid receptor agonists (SCRA), are increasingly smuggled into prisons via infused mail. Consumption of those substances by inmates in prisons is associated with increased aggression, violence, and organized crime. Onsite detection of infused mail often is challenging. Because the infused papers do not show any visible stains or olfactory changes, physical inspection is often insufficient. The applicability of further conventional on-site detection methods like immunoassays and sniffer dogs is severely limited. Because of the rapidly changing supply of already circulating and newly emerging NPS, it is impractical to impossible to keep up with the development of immunoassays or the training of sniffer dogs. Hence, confiscated mail samples are routinely tested by either liquid chromatography or gas chromatography coupled to mass spectrometry (MS), which is costly and time-consuming. In this study, recent advancements in the hyphenation of laser ablation (LA) and molecular MS were investigated regarding the possible application for the rapid and easy detection of NPS in prison mail. Utilizing an in-house developed LA-MS hyphenation based on atmospheric pressure chemical ionization (APCI), 31 mail samples confiscated in German prisons were analyzed. It was possible to correctly identify 27 samples containing SCRAs. For these positive samples, it was also possible to detect the specific compounds each paper was infused with. The use of LA-APCI-MS has simplified sample preparation and reduced analysis time per sample to 1 min.

Phenibut is a new psychoactive substance (NPS) first synthesized in Russia in 1963 as a derivative of gamma-aminobutyric acid. Originally developed for therapeutic use, it has gained popularity for nonmedical purposes, including recreational and cognitive enhancement. In Brazil, phenibut is uncontrolled and easily purchased online. This study used wastewater-based epidemiology (WBE) to investigate phenibut use patterns in two northeastern Brazilian cities. Composite daily wastewater samples were collected from two treatment plants (WWTPs), Recife (WWTP_A) and Olinda (WWTP_B), during two periods in 2023: Carnival and a reference week. Samples underwent solid-phase extraction (SPE) and analysis by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Phenibut concentrations were converted to population-normalized mass loads (PNMLs, mg/day/1000 inhabitants). The highest phenibut levels and PNMLs (up to 4.06 mg/day/1000 inhabitants) occurred during Carnival at WWTP_A, located in a major tourist area, suggesting recreational use. During the reference week, PNMLs ranged from detection limits to 2.29 mg/day/1000 inhabitants on weekdays, indicating possible functional or cognitive enhancement use. These findings reveal two distinct use patterns: recreational peaks during Carnival weekends and possible functional use on weekdays outside festive periods. This is the first evidence of phenibut detection in Brazilian wastewater and its temporal use patterns. The results highlight WBE's value in monitoring NPS trends and suggest recreational use predominates during large events. This underscores the need for public health attention and regulatory monitoring of uncontrolled substances with abuse potential.